Acylamino-cephalosporanic acids

ABSTRACT

Acylaminocephalosporanic acids of the formula I   wherein R1, R2 and R3 each represents hydrogen or lower alkyl radicals in which case R1 and R2 may also form together an optionally substituted alkylene radical, A represents an optionally substituted phenylene or thienyl radical, and X represents an oxygen atom or a single bond, the physiologically tolerable salts thereof, pharmaceutical preparations effective against bacterial infections containing these compounds, and a process for the manufacture.

United States Patent Schorr et al.

[ ACYLAMlNO-CEPHALOSPORANIC ACIDS [75] Inventors: Manfred Schorr, Frankfurt am Main; Elmar Schrinner, Wiesbaden, v both of Germany [73] Assignee: Farbwerke Hoechst Aktiengesellschaft vormals Meister Lucius & Bruning, Frankfurt/Main, Germany [22] Filed: Apr. 14, 1972 [21] Appl. No.: 244,287

[30] Foreign Application Priority Data Apr. 17, 1971 Germany 2118635 [52] U.S. Cl. 260/243 C, 424/246 [51] Int. Cl C07d 99/24 [58] Field of Search 260/243 C [56] References Cited UNITED STATES PATENTS 3,457,257 7/1969 Hanessian et a1. 260/243 C 3,484,437 12/1969 Urech et a1. 260/243 C 3,575,969 4/1971 Morin et al. 260/243 C 3,579,506 5/1971 Lemieux et a1. 260/243 C 3,632,578 1/1972 Chauvette 260/243 C 1 Feb. 11, 1975 3,708,477 l/l973 Martel et al. 260/243 C 3,749,711 7/1973 Schorr et a1. 260/239] Primary Examiner-Nicholas S. Rizzo Attorney, Agent. or Firm-Curtis, Morris & Safford [57] ABSTRACT Acylaminocephalosporanic acids of the formula 1 2 H -O-COCH R5 O r 2 3 coon 8 Claims, No Drawings ACYLAMINO-CEPHALOSPORANIC ACIDS 4-(2-imidazolinyl)-phenylacetic acid 4-( l ,4,5,(i-tetrahydro-Z-pyrimidyl)-phenylacetic acid 4-( l-methyll ,4,5,6-tetrahydro-2-pyrimidyl phenylacetic acid 4-[5,5-bis( 2-methoxyethyl )-l ,4,5,6-tetrahydro-2- pyrimidyl]-phenylacetic acid 4-( l ,S-dimethyl)-2-imidazolinyl)-phenylacetic acid 4-( l-methyl-5-butyl-2-imidazolinyl)-phenylacetic This invention provides acylamino-cephalosporanic 5 acid acids of the formula I 4-(-5,5-dimethyl-l ,4,5,6-tetrahydro-2-pyrimidyl)- Rl"N\c A y CH 00 NH-- S R -N 2 CH -O-GO-CH I v COOH wherein R, R and R each represents a hydrogen atom Phenylacetic or lower alkyl radicals, and R and R together may 15 'KY F form an alkylene radical optionally carrying substitucyclopemyllfupynfnldyl'z)'Phenylacetlc cats, A represents a phenylene or thienylene radicals optionally carrying substituents and X represents an Phenylacetlc acid Oxygen atom or a Sing|e bond 4-(2,4-diazaspiro[5,5]undec-2-en-3-yl)-phenylacetic This invention also provides a process for the manu- 2O acid facture of said acylamino-cephalosporanic acids of the P l l- -y )-P y formula I and the salts thereof which process comprises acld reacting 7-amino-cephalo-sporanic acid (7-ACS) or a 4'N,N"tetramethylene'amidino'phe"ylacetlc acid salt thereof with a carboxylic acid of the formula II 3'amldino'phenylacetic acid 4-amidino-2-methyl-phenylacetilc acid or R N 4-amidino-2'methoxy-phenylacetic acid C .A X. CH CQOH 1 4-amidino-2-butoxy-phenylacetic acid 2 2 4-amidino-2-chloro-phenylacetic acid or the corre- R N spondmg phenoxy-, thienylor thienyloxyacetic 3 acids.

a The carboxylic acids of the general formula ll are wherein 1 2 3 A and X are defined as above, esPeprepared in the known manner starting from 4-cyanocially in the form of a derivative capable of reacting P l y 'p lf' 5 cyanothlenylwith an amino group or f a 5a f such a compound cyanothlenyloxy-acetic acid esters. After conversion of If Rt, 2 and each represents an atkay] i l the cyano group into an imino ether the compounds are straight-chain or branched alkyl radicals carrying from reacted with ammonia amln'e or diamine to yield 1 to 5 carbon atoms can b used, h total number f amidine and the carboxylic acid ester group or an acid carbon atoms in radicals R, R and R preferably not amide group derived therefrom is saponified. cxcgeding The novel acyl derivatives of the 7-aminocephalospo- Suitable alkylene radicals are especially those con- 40 Panic acid are advantageously obtained by reacting a taining from 2 to 4 carbon atoms. As substituents for salt of the 7-aminocephalosporanic acid, for example, the alkylene radical there may be mentioned, for examan alkali metal or tertiary amine salt, such as the sople, low-molecular alkyl radicals having from I to 4 dium, potassium or triethylamine salt, with a reactive carbon atoms which may themselves be closed to a ring derivative of a carboxylic acid of the general formula optionally interrupted by a hetero atom, preferably an ll, if possible, at a neutral pH, preferably at a pH of oxygen atom. The alkyl substituents of the alkylene from 6 to 9. radical which are not closed to a ring, may also contain The salts of the 7-aminocephalosporanic acid may be a hetero atom, preferably an oxygen atom. used directly or may be prepared in the solution used A especially stands for the l,4-phenylene Or the for the reaction of 7-aminocephalosporanic acid and, thienylene radical which radicals may themselves be f l di hydrogencarbonate, di i h substituted, for example, by low-molecular alkyl, aldrogenphosphate or triethylamine.

I0 4 Carbon atoms) 0r halogen, Preferably y In general, the reaction in accordance with the invenchlorine. tion is carried out in water. It may also be worked in the For the process of the invention, the following compresence of solvents miscible with water, for example, pounds of formula II are suitable as starting materials: acetone, di th lf id di h l i di- 4-amidino-phenylacetic acid oxan or tetrahydrofuran.

4-N-methyl-amidino-p y acid The acylation of the 7-aminocephalosporanic acid in 4-NtN-dimethyl'amidino-phenylacetic acid accordance with the invention is suitably carried out 4N,N'dimethyl-amidino'phenylacetic acid with the acid chlorides of the carboxylic acids of the 4-N,N,N'-tflmethyl-amidino'phenylacefic acid general formula ll. They may be obtained in known 4"Nethyl-amidino'phenylacetic acid manner from the carboxylic acids by the action of thio- 4-N,N"dipropyl'amldino'phenylacetlc acid nyl chloride in an inert solvent, for example, an aro- -p y F acid matic hydrocarbon. They are obtained as hydrochlo- 4-N,N-dimethyl- 'e y -P rides which can be used directly for the further reaction. Besides the acid chlorides, there may also be used other reactive derivatives of the carboxylic acids of formula ll, such as, for example, acid bromides, activated esters, for example p-nitrophenyl ester, p-nitrophenylthio'ester or cyanmethyl ester, acid azides or symmetrical or mixed anhydrides.

The acylation of the 7-aminocephalosporanic acid is advantageously effected in such a manner that equimolar amounts or a small excess of a compound of formula II are added in the form of its reactive derivative to the solution of a salt of the 7-aminocephalosporanic acid in water or in a mixture of water and solvents miscible in water. I

It has proved to be especially advantageous to introduce the hydrochloride of the acid chloride in solid form. To bound the hydrochloric acid two equivalents of a base, for example sodium hydrogencarbonate or triethylamine are previously added to the solution. The reaction is effected at room temperature or at slightly elevated temperature, preferably within the range of from -5 to +5". The pH-value which is first slightly alkaline is reduced to about 7 in the course of the reac-. tion. It is then adjusted to about 5 with an acid, preferably with a dilute hydrochloric acid so that the acylamino-cephalosporanic acid containing an amidino group is present in the form of an inner salt. In some cases it precipitates in this form from the reaction solution directly as crystals and can be suction-filtered. if no crystallization occurs, the solution can also be evaporated to dryness to isolate the novel cephalosporin, which is advantageously carried out by using the method of lyophilizing.

Thus, a product is obtained which contains besides the cephalosporin also the salts obtained during the reaction. When the starting products had advantageously been used in molar radio the impurities mainly formed in the course of the reaction are the alkali metal or amine salts. When using sodium hydrogencarbonate for the preparation of the 7amino-cephalosporanic acid and an acid chloride, the crude cephalosporin contains, for example, only sodium chloride as essential ingredient. Due to the smooth course of the reaction, the cephalosporin/salt mixtures are already so pure that they can be used directly for therapeutical purposes. They may, however, be further worked to yield the pure cephalosporins.

So, for example, the cephalosporin/salt mixture obtained can be dissolved in water in a highly concentrated form. The cephalosporin is separated from this solution generally in crystalline form. By suctionfiltration and washing with ice water, it may be obtained in a form practically free from salts.

The novel acylaminocephalosporanic acids of the Formula 1 contain in the molecule-an optionally substituted amidino group and have therefore amphoteric character. They form inner salts and are easily soluble in water with a pH of about 5. In the lR-spectrum, these compounds show the absorption band at l,770 cm characteristics of the B-lactam ring. The products may also be characterized by thin-layer chromatography. The content can be determined in the usual manner by means of iodometry.

The products in accordance with the invention can be used in substance, in the form of their physiologically tolerable salts or optionally in the form of the crude products described above. As physiologically tolerable salt forming agents of this kind there may be mentioned, for example, hydrochloric acid, hydrobromic acid, amidosulfonic acid, citric acid, acetic acid and suitable bases, such as sodium hydroxide, ammonia or ethanol amine. The salts are obtained, for example, by mixing equivalent amounts in solution and vaporization.

The novel acylaminocephalosporanic acids have excellent antibacterial properties. The spectrum of activity is extended to Gram-positive and Gram-negative bacteria. In comparison with other cephalosporins used in therapy distinct differences can be observed with re gard to the sensitivity of the individual germ. In some cases the known cephalosporins are largely surpassed by the novel products containing an optionally substituted amidino group so that they have decisive therapeutical advantages in the treatment of infections resulting from those bacteria. Table I shows a comparison of several of the novel cephalosporins with a known one. The values indicated are to be understood as minimum inhibition concentrations evaluated with streptococci (seriological group A) in the series dilution test using l0 germs per tube.

Table 1 Minimum inhibition concentration in a/ml Germ: Streptococcous seriological group A.

Substance MTC 3 0.0078 b 0.0 I56 c 00078 d 0.0l25 e 0.0156 c 0.05

ephalothin a 7-(4-amidinophenoxyacetylamino)-cephalospo ranic acid b 7-[4-( l-methyl-l ,4,5,6-tetrahydro-2-pyrimidyl)- phenylacetylaminol-cephalosporanic acid 7-(4-amidinophenylacetylamino)-cephalosporanic acid d 7-[4-(5,5-dimethyl-l .4,5,6-tetrahydro-2- pyrimidyl)-phenoxyacetylamino]-cephalosporanic acid e 7-( 5-amidino-2-thienylacetylamino)-cephalosporanic acid Table 2 Chemotherapcutical test Test animal: mouse Infection: Streptococcous seriological group A.

Substance Subcutaneous treatment Per 05 Dosage Surviving animals Dosage Surviving animals unit on the unit on the a/kg 2nd day 10th day a/kg 2nd day lOth day a 3 x 62.5 4 4 3 x 2.5 4 4 3 X [.25 4 l b 3 x 500 4 4 3 x 25 4 4 3 X 250 2 2 3 X 5 4 3 C 3 X l() 4 4 3 X L 4 4 3 X 50 3 3 3 X 0.75 2 2 cephalo: 3 X 1500 4 4 3 X 50 4 4 thin 3 X 750 l l 3 X 25 3 3 The tables demonstrate the valuable properties and the superior effect of the novel cephalosporins used against special infections.

The products in accordance with the invention are valuable therapeutics which are very suitable for the treatment of bacterial infections. They can be applied as such or in the form of galenic preparations in combination with therapeutically suitable carriers and ex cepients, for example tragacanth, lactose, talc, agaragar, for example, in the form of tablets, dragees, capsules or of solutions or suspensions which contain the active substance in an amount of about 50 to 1,000 mg, preferably of from 100 500 mg. For parenteral administration suspensions or solutions in water are preferably used. Other physiologically tolerable organic solvents may also be added, such as, for example, ethanol, polyglycols, and solubilizers. A combination with other active substances can also be used.

The following Examples illustrate the invention:

EXAMPLES The novel cephalosporins described in the following Examples were characterized by the R value of the thin layer chromatogram, in which silica gel (Merck) was used as layer and a 0.1 N aqueous ammonium acetate solution as eluent. The development of the plates was effected by the action of iodine vapour. The Rf value of the 7-aminocephalosporanic acid was 0.75 under these conditions.

EXAMPLE 1 7-(4-Amidinophenylacetylamino)-cephalosporanic acid a. 5.35 g of 4-amidinophenyl acetic acid (melting point 295297C under decomposition) were suspended in 45 cc. of dry benzene. Two drops of dimethyl formamide and 17.9 g of thionyl chloride were added and the solution was heated under reflux for 1% hours. After cooling, the solid product was suction-filtered, washed with dry benzene and dried in vacuo. 6.9 g of 4-amidinophenyl acetic acid chloride-hydrochloride. were obtained; melting point: l74-l77C under decomposition.

b. 588 mg of sodium bicarbonate and 544 mg of 7 -aminocephalosporanic acid were dissolved in a mixture of 20 cc. of water and 2 cc. of acetone. At about 0C within 10 minutes, 513' mg of 4-amidinophenyl- EXAMPLE 2 7-(4-Amidinophenoxyacetylamino )-cephalosporanic acid 294 mg of sodium bicarbonate and 272.2 mg of 7-aminocephalosporanic acid were dissolved at 0C in .a mixture of 10 cc. of water and 1 cc. of acetone. Within a period of time of about 10 minutes 298 mg of 4-amidinophenoxyacetic acid chloridehydrochloride were introduced, the mixture was stirred at 0C for 45 minutes and was filtered. The clear filtrate was adjusted at pH 5.0 with 0.1 N hydrochloric acid and the solvent was eliminated by freeze-drying. The loose lyophilisate (420 mg) contained in addition to the novel cephalosporin the sodium chloride formed during the reaction. The content of 7-(4-amidinophenoxyacetylamino)- cephalosporanic acid evaluated by iodometry was 68 percent which corresponded to the theoretical R value of 0.49.

In an analogous manner there was obtained:

7-[4-( 5 ,S-dimethyl-l ,4,5,6-tetrahydro-2-pyrimidyl phenoxy-acetylamino]-cephalosporanic acid; R,- value 0.22.

7-[4-(9-oxa-2,4-diazaspiro[5,5] undec-2en 3-yl)- phenoxy-acetylaminol-cephalosporanic acid; R,- value 0.18.

7-[4-( l -methyl-l ,4,5,6-tetrahydro-2-pyrimidyl)- phenylacetylamino] -cephalosporanic acid; R,- value 0.18.

.7-[4-( 2-imidazolinyl)-phenoxyacetylamino]-cephalosporanic acid; R value 0.41.

What is claimed is: 5. A compound as claimed in claim 1, wherein R and 1. An acylaminocephalosporanic acid of the formula R2 together Stand for p py a is y A O N CH2OC0CH3 and the physiologically tolerable salts thereof, wherein resents p-phenylene and X is a single bond. R and R taken alone, are hydrogen or alkyl having 1 6. A compound as claimed in claim 1, wherein R and to 5 carbon atoms or, taken together, are alkylene hav- 2 together Stand for ing 2 to 4 carbon atoms which may be substituted by Cur MGMCH, alkyl having 1 to 4 carbon atoms which may contain an oxygen atom or by alkyl substituents having I to 4 car- CH (ll-l bon atoms closed to a ring which may be interrupted by an oxygen atom; R is hydrogen or alkyl having 1 to 5 R3 A represents P'Phen y|ene and x qxygem carbon atoms; A is phenylene or thienyl which may be A Compound as Claimed Clam 1, Where!" 1 and substituted by alkyl having 1 t0 4 carbon atoms, alkoxy 2 together standlfol' of l to 4 carbon atoms, or halogen; and X is an oxygen atom or a single bond.

2. A compound as claimed in claim 1,.wherein R R and R each stand for hydrogen, A represents pphenylene and X is a single bond. 2

3. A compound as claimed in claim 1, wherein R1, R and R each stand for hydrogen, A represents p- 3 R3 is hydrogen,

Phenylene and X is g 0 A represents p-phenylene and X is oxygen.

A compound as clamed m clam Wherem R R2 8. A compound as claimed in claim 1, wherein R and and R3 each Stand for f F A :epresems 'Fh R together stand for ethylene, R is hydrogen, A repreand amidino radical bemg hnked in the 5-position of Sems p phenylene and X is oxygen the thienyl radical, and X is a single bond. 

1. AN ACYLAMINOCEPHALOSPORANIC ACID OF THE FORMULA
 2. A compound as claimed in claim 1, wherein R1, R2 and R3 each stand for hydrogen, A represents p-phenylene and X is a single bond.
 3. A compound as claimed in claim 1, wherein R1, R2 and R3 each stand for hydrogen, A represents p-phenylene and X is oxygen.
 4. A compound as claimed in claim 1, wherein R1, R2 and R3 each stand for hydrogen, A repreSents 2-thienyl, and amidino radical being linked in the 5-position of the thienyl radical, and X is a single bond.
 5. A compound as claimed in claim 1, wherein R1 and R2 together stand for n-propylene, R3 is methyl, A represents p-phenylene and X is a single bond.
 6. A compound as claimed in claim 1, wherein R1 and R2 together stand for
 7. A compound as claimed in claim 1, wherein R1 and R2 together stand for
 8. A compound as claimed in claim 1, wherein R1 and R2 together stand for ethylene, R3 is hydrogen, A represents p-phenylene and X is oxygen. 